THROMBIN INHIBITORS, THE PREPARATION THEREOF AND THE USE THEREOF FOR THERAPEUTICAL, PROPHYLACTIC AND DIAGNOSTIC APPLICATIONS

Abstract: Peptides having 25 to 27 amino acids capable of binding both to the catalytic site and to the non-catalytic site of hirudin.

Description: THROMBIN INHIBITORS, THE PREPARATION THEREOF AND THE USE THEREOF FOR THERAPEUTICAL, PROPHYLACTIC AND DIAGNOSTIC APPLICATIONS The present invention relates to thrombin inhibitors, the preparation thereof and the use thereof for therapeutical, prophylactic and diagnostic applications. Thrombin is a serin-protease involved in various important physiological functions [Fenton, J.W. (1981) Annal. N.Y. Acad. Sci., 370, 468-495; Bar Shavit, R. and Wilner, G.D. (1986) Int. Rev. Exp. Pathol., 29, 213-241]. Besides playing a role in blood coagulation, thrombin also has other activities, among which a strong mitogenic signal with fibroblasts in vitro, and it also exerts a chemotactic effect on monocytes (Bar Shavit). Thrombin stimulates the formation of cGMP in neuroblastoma cells [Snider, R.M. et al. (1984), J. Biol. Chem. , 259, 9078-9081; Snider, R.M. (1986), Ann. N.Y. Acad. Sci., 485, 310-313]: it is involved in the control of neuronal differentiation [Gurwitz D. and Cunningham, D.D. (1988), Proc. Natl. Acad. Sci., 85, 3440-3444], Thrombin seems to also have a role in promoting cancer, due to the capability of fibrin, which is its digestion product, to act as a substrate for tumour growth [Falanga A. et al. (1985) Biochemistry, 24, 5558-67; Gordon, S.G. et al. (1985), Blood, 66,1261-65; Falanga A. et al . , (1988), Blood, 71, 870-75]. Many pathologies being a high risk for health, such as cardiac infarction, thrombosis, peripheral arteries occlusions, and the like, require a regulation of thrombin activity. The physiological inhibitor of thrombin in blood is antithrombin III, but thrombin is poorly inhibited by it [Rosenberg, R.D. (1977), Fed. Proc, Fed. Am. Soc. Exp. Biol., 36, 10-18]. Heparin increases thrombin activation rate by at least 3-4 magnitudes [Olson, S.T. and Shore, J.D. (1982), J. Biol. Chem., 257, 14891- 14895], therefore it is commonly used as a therapeutical agent, e.g. in venous thromboembolism, in which thrombin activity is responsible for the development or the expansion of a thrombus. However, the use of heparin has some drawbacks: it is ineffective in patients lacking antithrombin III and it has a short half-life in blood; it fails to interrupt platelet-mediated arterial thrombosis or the formation of the hemostatic plug [Salzman, E.W. et al. (1980), J. Clin. Invest., 65, 64-73; Hanson, R.S. and Harker, L.A. (1988), Proc. Natl. Acad. Sci., USA, 85, 3184-3188]. Due to these and other drawbacks, a search for different compounds useful for a fast inactivation of thrombin has been required. Hirudin, a polypeptide of molecular weight 6950 excreted by the salivary glands of Hirudo medicinalis, belongs to a family of polypeptides containing about 65 single chain amino acids (a.a.) and it is a strong specific thrombin inhibitor [Chang, J.Y. (1983), FEBS, 164 (2), 307-313; Konno, S. , et al., G.B. 1988, Arch. Bioch. Biophys. 267, 158-166; Fenton J.W. et al. (1988), Biochemistry, 27, 7106-7112; Stone, S.R. et al (1986), Biochemistry, 25, 4622-4628]. Hirudin binds t thrombin (K.--6.3 x 10" 1 M) forming a non covalen complex 1:1; such an interaction occurs at a differen site from the enzyme catalytic site, even thoug hirudin additionally also binds to the active site Therefore, in hirudin itself, three different domain can be evidenced: i) the N-terminal segment penetratin into thrombin catalytic site; ii) the C-termina segment, of about 18 residues, which binds in a mainl cationic groove of thrombin; iii) the central nucleu (residues 5-48) acting as a spacer of the two N- and C terminal domains. Hirudin has successfully been used as a anticoagulant and antithrombotic agent. Contrary t heparin, hirudin requires no presence of endogenou cofactors such as antithrombin III and it is not boun or inactivated by platelet factors or by othe substances having antiheparin effects. Hirudin differ from heparin in that it causes no bleeding side effects. Notwithstanding said advantages and even thoug hirudin is nowadays prepared by recombinant DN techniques [Markwardt, F. (1991), Haemostasis, 21 Suppl. 1, 11-26], the clinical use of hirudin i restricted mainly since the substance is not availabl in sufficient amounts for therapeutical purposes. Therefore a number of researchers are involved i efforts to synthesize hirudin peptide fragments whic are easier and less expensive to prepare and have an antithrombotic activity similar or even higher tha that of hirudin. Compounds inhibiting thrombin are known i literature and among them particularly interesting ar hirudin peptide fragments [EP-468448; WO-9119734; GB 2242681; EP-443598; EP-443429; EP-421367; WO-9101328 WO-9101142; EP-372670; EP-372503; EP-364942; EP-341607 9049 US-4971953; EP-333356; EP-291982; EP-276014; WO 9102750; J.M.Maraganore, et al., Biochemistry, (1990) 29, 70957101; J.M.Maraganore, et al., J. Biol. Chem. (1989) , 264, 86928698; M.Scharf, et al., FEBS, (1989) 255, 105110), W.J.Hoekstra, et al., Tetrahedron (1992), 48, 207318; P.J.Braun, et al., Biochemistry (1988), 27, 65176522; A.Wallace, et al., Biochemistry (1989), 28, 1007910084; J.Dodt, et al., FEB, (1988) 229, 8790; T.J.Owen, et al., J. Medicinal Chem. (1988), 10091011; J.L.Krstenansky, et al., J. Medicina Chem., (1987), 16881691; J.B.Lazar, et al. J.Biol.Chem. , (1991), 266, 685688; S.Dennis, et al. Eur.J.Biochem. , (1990), 188, 6166; V.Steiner, et al. Biochemistry, (1992), 31, 22942298; A.Betz, et al. Biochemistry, (1992), 31, 45574562, S.J.Di Maio, e al., J.Biol.Chem. , (1990), 265, 2169821703]. Among them, two class of compounds ar particularly relevant: i) those reproducing the C terminal domain of hirudin (48-65 or smaller sizes which therefore bind only to the non-catalytic site o thrombin; ii) those which, analogously to hirudin contain an N-terminal domain penetrating into th catalytic site, a spacer and a domain corresponding t the C-terminal segment of hirudin which binds to th non-catalytic site. The latter compounds penetrate in the catalytic site of thrombin thereby forming a β strand which is antiparallel with the Ser 214 -Gly 2 -'- residues such as in the case of other serineprotease inhibitor complexes and therefore they can be subjecte to hydrolysis [E.SkrzypczakJankun, et al. , J.Mol.Biol (1991), 221, 13791393; I. .Witting, et al., Biochem.J (1992), 283, 737-743]. The present invention relates to novel hirudi analogues, for which the name "hirunorms" is proposed with a chemical composition different from the up t now developed ones. Said compounds, showing an overal biological activity higher than that of already know products, have the following general formula (I): G1-G2-G3-G4-G5-G6-G7-G8-G9-F1-F2-F3-F4-F5-F6-E1-E2-E3- E4-E5-E6-E7-E8-E9-E10 wherein: Gl - lie, Val, Leu, Cha, Chg, alle, Nle, Pro, Pip, Phe Tyr, Trp, Pgl, 1-Nal, 2-Nal, Met G2 - Arg, Val, L-Pap, L-Map, Ala, lie, Leu, Lys , Orn Thr, aThr, alle, Ser G3 - 2-Nal, Phe, 1-Nal, Trp, Tyr, Cha, Pgl, Chg G4 - Thr, Ser, Asn, Gin, His, aThr G5 - Asp, Glu G6 - D-Ala, Aib, Gly, Ac 6 c, ACrC G7 - -Aca, g-Ava, β-Ala-β-Ala, β-Ala-β-Ac 5 c, β-Ala-β Ac 6 c, β-Ala-Gly, β-Ala-Gaba, Gly-β-Ala, Gly-Gly Gly-Gaba, Gly-^-Ava, Gaba-Gly, Gaba-β-Ala, £-Ava Gly, Gly-Gly-Gly G8 - Pro, Ala, Gly, Ac 6 c, Ac 5 c G9 - Glu Fl - Ser, Asn, Gin, Thr, aThr, His, h-Ser, h-Phe F2 - His F3 - h-Phe, Leu, Asp, Asn, Gly, Nle, lie, alle, Val Cha, Chg, Phe, Tyr, Pgl, Trp, 1-Nal, 2-Nal, Met Deg, Dpg F4 - Gly, Asp, Asn F5 - Gly, Ala, D-Ala, Deg, Dpg F6 - Asp El - Tyr, Phe, Trp, Cha, His, Pgl, 1-Nal, 2-Nal, Chg E2 - Glu E3 Glu, Pro, Sar, Hyp,Λ-Pro, thioproline, Pip, Azt E4 - lie, Chg, Cha, E5 - Pro, Sar, Hyp, Δ-Pro, thioproline, Pip, Azt E6 - Aib, Ala, Ac g c, Ac 5 c, Ac 4 c, Ac 3 c, cis or transΔ- Glu, (S) or (R)-OMe-Glu, Asp, Glu E7 - Aib, Ala, Ac g c, Ac 5 c, Ac 4 c, Ac 3 c, cis or transΔ- Glu, (S) or (R)-^Me-Glu, Asp, Glu E8 - Tyr, cis or trans -Tyr, (S) or (R)-O(Me-Tyr, Phe, Aib, Ala, para or meta N0 2 _Phe, para or met SO^H-Phe, para or meta P0 3 H 2 -Phe, para or met P0 4 H 2 -Phe, para or meta S0 4 H-Phe, meta S0 3 H-Tyr meta P0 3 H 2 -Tyr, para or meta CH 2 P0 3 H 2 -Phe, para o meta CH 2 so 3 H-Phe, para or meta CH 2 COOH-Phe, par or meta CO,H-Phe E9 - Cha, Leu, cis or transΔ-Leu, (S) or (R) (VMe-Leu ElO -D-Glu, Glu, β-Ala, Asp, D-Asp, Gly, Ala, Gin, Gla Particularly preferred are: a) compounds of general formula (I) wherein: Gl - lie or Val or Leu or Cha or Chg or alle or Nle o Pro or Phe or Tyr or Trp or Pgl or 1-Nal or 2-Na or Met; G2 - Arg or L-Pap or L-Map or Val or Th or He or Ala or Ser; G3 - 2-Nal or Phe or 1-Na or Trp or Tyr; G4 - Thr; G5 - Asp; G6 - D-Ala G7 - Gly-β-Ala or β-Ala-β-Ala; G8 - Pro; G9 - Glu Fl - Ser or Asn or h-Ser or h-Phe; F2 - His; F3 Nle or Cha or Met or h-Phe; F4 - Gly; F5 « Gly F6 - Asp; El - Tyr or Phe; E2 - Glu; E3 - Glu o Pro; E4 - He; E5 - Pro; E6 - Aib; E7 - Aib E8 - Tyr or P0 4 H 2 -Phe; E9 = Cha or Leu; ElO - D Glu or Glu; b) compounds of formula (I) wherein: Gl - Chg or He or Val or alle or Nle or Phe or Tyr o Trp or Pgl or 1-Nal or 2-Nal; G2 - Arg or Val ; G - 2-Nal or Phe or 1-Nal; G4 - Thr; G5 - Asp G6 - D-Ala; G7 - Gly-β-Ala; G8 = Pro; G9 - Glu Fl - Ser; F2 - His; F3 - Nle or h-Phe; F4 - Gly F5 - Gly; F6 - Asp; El - Tyr or Phe; E2 = Glu E3 - Glu or Pro; E4 - He; E5 - Pro; E6 - Aib E7 - Aib; E8 - Tyr or P0 4 H -Phe; E9 = Cha or Leu ElO - D-Glu or Glu. In the following, the abbreviations used in th disclosure are reported: Orn - ornithine, Gly - glycine, Ala- alanine, Va = valine, Leu « leucine,Δ-Leu - dehydroleucine, 0 Me- Leu - (X -methylleucine, He - isoleucine, Pro - proline Phe = phenylalanine, Trp = tryptophan, Met ethionine, Ser - serine, Thr = threonine, Tyr tyrosine, Δ-Tyr -Q -β-dehydrotyrosine, 0^ Me-Tyr -(X- methyltyrosine, Asn - asparagine, Gin -= glutamine, As = aspartic acid, Lys - lysine, His - histidine Glu - glutamic acid, Arg - arginine, Nle - norleucine Hyp - hydroxyproline, Δ-Pro - dehydroproline, Λ-Glu - (V-β-dehydroglutamic acid, (YMe-Glu - (Y-methyl-glutamic acid, Pgl «- phenylglycine, 1-Nal - β-1-naphthylalanine, 2-Nal - β-2-naphthylalanine, Cha = cyclohexylalanine, alle - allo-isoleucine, Chg - cyclohexylglycine, h-Phe - omophenylalanine, h-Ser = omoserine, Sar - sarcosine, Pip - pipecolic acid, Azt - azetidinic acid, gla - Y- carboxyglutamic, Pap - para-amidino-phenylalanine, Map - meta-amidino-phenylalanine, Deg - diethylglycine, Dpg - dipropylglycine, aThr - allo-threonine, Aba - C - amino-n-butyric acid, Pba - Q(aminophenylbutyric acid, Aib- (Y-aminoisobutyric acid (O^-methyl-alanine) , Dap - 2,3 diaminopropionic acid, Dab = 2,4 diaminobutyric acid, Gaba - -aminobutyric acid, £ ^ Aca - -aminocaproic acid, C-Ava - -aminovaleric acid, β-Ala - β-alanine, Ac,c = (^-aminocyclopropanecarboxylic acid, Ac.c = 0(~ aminocyclobutanecarboxylic acid, Ac-c = θ(~ am i noc y c l°~ pentanecarboxylic acid, Ac g c = (X -aminocyclohexanecar- boxylic acid, β-Ac,-c = β-aminocyclopentanecarboxylic acid, β-Ac fi c = β-aminocyclohexanecarboxylic acid, NO ? - Phe - nitrophenylalanine, S0 3 H-Phe - phenylalanine sulfonate, PO,H 2 -Phe = phenylalanine phosphonate, PO.H 2 -Phe - phenylalanine phosphate, SO.H-Phe = pheny¬ lalanine sulfate, S0 3 H-Tyr = tyrosine-m-sulfonate, P0 3 H 2 -Tyr = tyrosine- -phosphonate, CH 2 P0 3 H 2 -Phe = phenylalanine methylphosphonate, CH 2 S0 3 H-Phe = phenylalanine methylsulfonate, CH 2 C00H-Phe = phenylala¬ nine acetate, C03H-Phe - phenylalanine carbonate, Boc - tert-butyloxycarbonyl, Firtoc - fluorenylmethoxycarbonyl, Bzl - benzyl ester, PAM - phenylacetoxymethyl, TFA - trifluoroacetic acid, DCM - dichloromethane, DIEA - diisopropylethylamine, DMF - dimethylformamide, OBzl - benzyl ether, PyBop = benzotriazole-1-yl-oxy-tris- pyrrolidino-phosphonium-hexafluorophosphate, DCC dicyclohexylcarbodiimmide, DCLT - dicyclohexylurea, Bom - -benzyloxymethyl, Tos - tosyl, BSA - bovine seroalbumin. As far as the definitions of the different amino acids are concerned, reference is made to The Biochemical Journal 219, N β 2, 345-373 (1984). The peptides of the present invention contain a number of charged groups on the amino acids side chains and of course they have to be neutralized with suitable counter-ions. Since any positively charged chemical group can be a potential counter-ion, only the pharmaceutically acceptable ones will be used, i.e. those cations which are non toxic at the used doses and do not have in their turn any undesired actions. Non- limiting examples of counter-ions are: alkali or alkaline-earth metal ions, or Al 3+ , primary, secondary and tertiary organic amines. The peptides of the present invention can of course be administered also in the acidic form by adding inorganic or organic acids, such as HCl, HBr, H 2 SC>4 , H PC> or acetic, piruvic, succinic, glutaric, fumaric, malic, tartaric, citric, ascorbic, maleic acid. The peptides of the present invention can also be prepared by the various techniques reported in literature, see e.g. Schroeder et al. "The Peptides" vol. 1, Academic Press, 1965; Bodanszky et al. "Peptide Synthesis Interscience Publisher, 1966; Barany & Merrifield, "The peptides; Analysis, Synthesis, Biology", 2, Chapter 1, Academic Press, 1980. Said techniques include peptide synthesis in solid phase, peptide synthesis in solution, organic chemistry synthetic methods, or any combination thereof. The selected synthesis scheme will depend of course on the composition of the particular molecule. Preferably, the claimed molecules being entirely on a peptide basis, synthetic methods based on suitable combinations of solid phase technique and conventional solution methods are used, which involve low costs particularly on industrial scale. The synthesis of peptides containing sulfonated, sulfated, phosphated or phosphonated tyrosines, or containing methylphosphonated or methylsulfonated phenylalanines, can be carried out according to one of the methods described in literature [J.M. Lacombe et al., Int. J. Peptide Protein Res., (1990), 36, 275-280; C. Garbay-Jaureguiberry et al., Int. J. Peptides Protein Res., (1992), 39, 523-527]. Preferably, the synthesis of the peptides containing aromatic amino acids substituted with anion groups at one of the meta or para positions of the ring, can be carried out by means of two alternative procedures: i) "Global Approach"; ii) "Building Block Strategy". The first procedure involves the complete synthesis, both with Boc and F oc chemistry, of the desired peptide, on which the residues to be phosphorylated with the side chains unprotected are introduced. The phosphorylation reaction is then carried out using different reagents and a variety of phosphite-protecting groups: (isopropyl) 2 N-P(OR) 2 wit R-ethyl, benzyl, methyl, tert-butyl. The subsequen oxidation of the phosphite by tert-butyl peroxide an the cleavage from the resin yields the desire phosphopeptide. The second strategy makes use of the previousl phosphated tyrosine, which can be introduced by mean of the solid phase technique, with both Boc and Fmo chemistry. The compounds of the present invention (hirunorms contain three domains (as the hirudin natura molecule): 1) a C-terminal domain; 2) a spacer arm; 3 an N-terminal domain. It should be pointed out that for the used structural solutions, the hirunorm reproduce the action mechanism of hirudin. Hirudin interacts with thrombin on two differen regions: a groove different from the catalytic site being mainly cationic, and the catalytic site itself The N-terminal segment of hirudin penetrates into thi site, thereby forming a β-strand parallel with th Ser 214 Gly 21 residue, and therefore it cannot b hydrolyzed by thrombin itself. The C-terminal segmen interacts with a number of amino acid residues in th groove far from the catalytic site. Hirudin centra nucleus has poor interactions with thrombin itself, bu it plays an important role in the parallel location o the N-terminal segment of hirudin in the catalytic sit [T.J.Rydel, A.Tulinsky, .Bode, R.Huber, J.Mol.Biol. (1991), 221, 583601]. A schematic comparative analysis of th structural, functional and biological characteristic of the compounds of the invention compared with hirudi is reported below. 1) The compounds of the invention have molecular weight, at least the half of that o hirudin, therefore requiring synthetic procedure easier on industrial scale. 2) The N-terminal moiety has a compositio similar to the one of hirudin and it has access to th catalytic site from the same direction, thus it is no subjected to the amidolytic action of thrombin itself. 3) The linker moiety is at least 3 times smalle than hirudin and the optimization of the interaction related to the use of particular non protein amin acids, makes more specific the interaction wit thrombin. 4) In the C-terminal moiety, more than 40% o the amino acidic residues are different from those o hirudin and they contribute in optimizing th interactions between this peptide segment and th cationic groove on thrombin surface. Hereinafter, the peptide sequences of the example of the invention are reported. a) hirudin; b) hirunorm-I; c) hirunorm-II; d hirunor -III; e) hirunorm IV; f) hirunorm V. A) The numeration refers to hirudin amino aci residues; residues 6 to 47 are not reported. I) N Terminal moiety; II) linker moiety; III) C-termina moiety. The amino acid substitutions never used befor to prepare hirudin-related peptide analogues ar indicated in bold-type. Tyr(-) means phenylalanine phosphate. 144 13 A a b c d e f 1 He He He He Chg Chg I) 2 Val Arg Arg Arg Arg Val 3 Tyr Tyr Phe Phe 2-Nal 2-Nal 4 Thr Thr Thr Thr Thr Thr 5 Asp Asp Asp Asp Asp Asp * * D-Ala D-Ala D-Ala D-Ala D-Ala * * β -Ala β-Ala β-Ala β-Ala β-Ala 48 Pro Pro Pro Pro Pro Pro 49 Glu Glu Glu Glu Glu Glu 50 Ser Asn Ser Ser Ser Ser II) 51 His His His His His His 52 Asn Asn h-Phe h-Phe h-Phe h-Phe 53 Asx Asn Gly Gly Gly Gly 54 Gly Gly Gly Gly Gly Gly 55 Asp Asp Asp Asp Asp Asp 56 Phe Phe Tyr Tyr Tyr Tyr 57 Glu Glu Glu Glu Glu Glu 58 Glu Glu Glu Glu Glu Glu 59 He He He He He He 60 Pro Pro Pro Pro Pro Pro III) 61 Glu Aib Aib Aib Aib Aib 62 Glu Aib Aib Aib Aib Aib 63 Tyr Tyr Tyr yr(-) Tyr Tyr 64 Leu Leu Cha Cha Cha Cha 65 Gin Glu D-Glu D-Glu D-Glu D-Glu In conclusion, the claimed peptides have th following characteristics: 1) They have been designed to function analogously t hirudin and differently from the other peptide inhibitors analogues of hirudin. 2) They show activities higher than hirudin. 3) They are synthetically simpler than hirudin. However, it is necessary to stress how, due to the particular kind of interaction of the claimed molecules with thrombin, other amino acid substitutions, non specifically envisaged but within the peculiar molecular structure of hirunorms, can be carried out to improve the biological strength. Accordingly, owing to the structural and functional characteristics thereof, the peptides of the invention proved to be stronger specific inhibitors of thrombin. The efficacy of hirunorms was evaluated in vitro by measuring APTT (Activated Partial Thromboplastin Time), PT (Prothrombin Time) and TT (Thrombin Time) in the presence of increasing amounts of the product on healthy donors' plasma and compared with normal values and values obtained with hirudin. The efficacy of the claimed molecules in inhibiting platelet aggregation was evaluated also in vitro using PRP (platelet-rich plasma of healthy donors) or PPP (platelet poor plasma) . The resistance of the compounds of the present invention versus plasmatic proteases was evaluated in vitro in two groups of tests. In the first one, the resistance to proteolysis of thrombin itself was tested incubating the claimed compounds in the presence o thrombin and analysing the mixture at different times In the second one, the claimed compounds were incubate in plasma of healthy volunteers and, after filtratio of the plasma proteins, the supernatants were analyze by HPLC and capillary electrophoresis. Moreover, the efficacy of the products of th present invention was also evaluated determining th inhibition constants IC versus thrombin, usin chromogenic substrates. The anticoagulant dose of a peptide of th invention can vary from 0.05 mg/Kg to 250 mg/Kg patien body weight per day, depending on the patient, th severity of the thrombotic condition and the selecte peptide. The term "patient" as herein used include mammals, such as primates, man included, ovines, bovines, horses, cats, dogs, pigs, rats, mice. Th suitable dose can be determined easily, preferably th treatment should envisage up to 4 daily doses which ca vary from 1 to 100 mg of active compound per dose. These compounds have a strong anticoagulant activity, therefore they can be used in the treatment an prevention of a wide variety of thrombotic conditions, particularly cerebro-vascular and coronary arteria diseases, more generally of all of those pathologies i which an anticoagulant action is beneficial. Th compounds can be used in combinations, compositions an methods for the treatment of cardiac infarction pulmonary embolism, venous thrombosis, periphera arteries occlusion, restenosis following arteria damage or invasive cardiologic surgery. They can also be used in the therapy of thromboti pathologies in combination with a thrombolytic agent t decrease the reperfusion time and prevent an reocclusions. In such a way, the dosage of the use thrombolytic agent can be decreased, thus preventin the involved side-effects, such as the risk o bleeding. The products of the invention can be coated i layered on the surface of invasive prostheses such a artificial valves, vascular grafts, catheters and th like, to prevent any formations of clots or aggregatio and platelet activation in patients carrying sai prostheses. The compounds of the present invention can also b used in preventing venous and arterial thrombosis an disseminated intravascular coagulation. Moreover, the can be used in the prophylaxis of arterial thrombosis particularly in cardiosurgery, to prevent coronary by pass occlusion and thrombotic reocclusion followin transluminal percutaneous coronary angioplasty, o after thro bolysis in venous and arterial vessels. The can also be used in extracorporeal circulation particularly in hemodialysis. The compounds of the invention can, besides in th prevention of vascular pathologies, be used in th prophylaxis of inflammatory responses, tumou pathologies and neuro-vegetative disorders, therefor the use thereof is important in the treatment o chronic or acute aterosclerosis, oedema an inflammations, tumours and metastases, an neurodegenerative diseases such as Parkinson's diseas and Alzheimer disease. The compounds of the present invention ar suitable for the therapeutical administration to highe animals and man through the parenteral, subcutaneous topical or nasal routes, attaining pharmacologica effects according to the above described properties Suitable forms for the oral administration are aqueou or oily solutions or suspension, emulsions, syrups elixirs or freeze-dried forms. Topical administratio can be effected by means of preparations such a aqueous gels, oily suspensions or emulsions. The dose of active ingredient in said compositions can rang from 0.1 to 10 g/kg body weight. The compounds of th invention can also be administered in form of depo injection or implant preparation, which can b formulated to allow a substantial release of the activ ingredient. The active ingredient can be compressed i tablets or small cylinders and subcutaneously o intramuscularly implanted in form of depot injection or implants. The implants can consist of iner materials, such as biodegradable polymers or syntheti silicons. The compounds of the invention can have a numbe of applications in diagnostics. They are indeed versatile tool to selectively prevent or interrup thrombin action in biochemical and diagnostic systems. The compounds can be used in kinetics studies o the fibrin-peptide release thrombin-catalyzed, of th fibrin structure and of the interaction betwee fibrinogen fragments and fibrin mono- and oligomers. The compounds can be used in studies of thrombi binding to cell receptors of platelet, endothelia cells, fibroblasts and cancer cells. The compounds can be added in an excess to blood, plasma or cell mixtures to prevent thrombin actio immediately after its production; they can be used t titre a thrombin specific amount. The compounds can be used to discriminate th activity of thrombin, of the precursors and cofactors thereof, and the activities of other plasma proteases. For the specific applications in diagnostics, the peptides of the invention can be, for example, the essential constituent of a kit for the determination of the concentration of factor IXa, of factor Xa of thrombin or of mixtures thereof in a biological sample. The peptides of the invention can also be labelled with a radioisotope for ex vivo imaging of fibrin or platelet thrombi. The following examples further illustrate the invention. EXAMPLE 1 Preparation of the compound hirunorm-I having structure He-Arg-Tyr-Thr-Asp-D-Ala-β-Ala-β-Ala-Pro-Glu-Asn- His-Asn-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Aib-Aib-Tyr- Leu-Glu-OH [compounds of formula (I) wherein Gl - He, G2 - Arg, G3 - Tyr, G4 - Thr, G5 - Asp, G6 = D-Ala, G7 - β-Ala-β-Ala, G8 - Pro, G9 - Glu, Fl - Asn, F2 - His, F3 - Asn, F4 - Asn, F5 - Gly, F6 - Asp, El - Phe, E2 - Glu, E3 - Glu, E4 - He, E5 - Pro, E6 - Aib, E7 - Aib, E8 - Tyr, E9 - Leu, ElO - Glu], corresponding to Sequence Id no. 1. An automatic peptide synthesizer was used for the preparation, starting from 0.714 g of Boc-Glu( f Bzl)- OCH2-PAM resin (0.70 meq/g) , equivalent to 0.5 mmol of amino groups. The various amino acids, suitably protected, are then reacted in the correct order. The acylation time is 1 hour. For the acylation with Boc- Aib-OH, the reaction mixture is added with PyBop (2 mmol) in DMF and coupling is repeated for 2 hours. All the other amino acids are coupled as symmetric anhydrides, dissolving 2 mmol of amino acid in 5 ml of DCM, the solution is cooled to 0°C and 1 ml of a 0.5 M DCC solution in DCM is added ; after 15 min DCU is filtered and the resulting solution is added to the deprotected resin. The dry resin is placed into a teflon reactor with 1 ml of anisole; the mixture is cooled to -50°C and 10 ml of hydrofluoric acid are distilled therein, then the mixture is stirred for 1 h in an ice bath. The product is dried, washed with ethyl ether (2 x 15 ml) and extracted with 50% acetic acid (3 x 15 ml), filtered through a porous filter to remove the exhausted resin. The resulting product is diluted with water and freeze-dried. The peptide is finally purified by reverse phase chromatography and characterized by analytic HPLC with a Vydac C18 0.46 x 25 cm column, with a linear gradient in acetonitrile containing 0.1% (v/v) of trifluoroacetic acid (phase B) against 0.1% (v/v) aqueous trifluoroacetic acid (Phase A) , from 5 to 70% in B in 35 min at flow rate of 1 ml/min, with UV detector at 210 nm. Retention time (Rt) - 24.8 min; chromatographic purity > 99%. EXAMPLE 2 Analogously, the following peptides are prepared: hirunorm-II of structure Ile-Arg-Phe-Thr-Asp-D Ala-Gly-β-Ala-Pro-Glu-Ser-His-h-Phe-Gly-Gly-Asp-Tyr- Glu-Glu-Ile-Pro-Aib-Aib-Tyr-Cha-D-Glu-OH [compounds o formula (I) wherein Gl = He, G2 - Arg, G3 - Phe, G4 Thr, G5 - Asp, G6 - D-Ala, G7 - Gly-β-Ala, G8 = Pro, G - Glu, Fl - Ser, F2 - His, F3 - h-Phe, F4 = Gly, F5 Gly, F6 - Asp, El - Tyr, E2 - Glu, E3 « Glu, E4 = He E5 - Pro, E6 - Aib, E7 - Aib, E8 - Tyr, E9 - Cha, ElO D-Glu]. Rt- 28.7 min (Sequence Id no. 2). hirunorm-IV of structure Chg-Arg-2-Nal-Thr-Asp-D Ala-Gly-β-Ala-Pro-Glu-Ser-His-h-Phe-Gly-Gly-Asp-Tyr- Glu-Glu-He-Pro-Aib-Aib-Tyr-Cha-D-Glu-OH [compounds o formula (I) wherein Gl = Chg, G2 = Arg, G3 = 2-Nal, G→ = Thr, G5 - Asp, G6 = D-Ala, G7 - Gly-β-Ala, G8 - Pro G9 - Glu, Fl - Ser, F2 = His, F3 - h-Phe, F4 - Gly, F = Gly, F6 = Asp, El «- Tyr, E2 = Glu, E3 = Glu, E4 He, E5 - Pro, E6 = Aib, E7 = Aib, E8 - Tyr, E9 - Cha ElO = D-Glu]. Rt- 21.1 min (Sequence Id no. 3). hirunorm-V of structure Chg-Val-2-Nal-Thr-Asp-D Ala-Gly-β-Ala-Pro-Glu-Ser-His-h-Phe-Gly-Gly-Asp-Tyr- Glu-Glu-He-Pro-Aib-Aib-Tyr-Cha-D-Glu-OH [compounds o formula (I) wherein Gl - Chg, G2 = Val, G3 - 2-Nal, G - Thr, G5 = Asp, G6 = D-Ala, G7 - Gly-β-Ala, G8 - Pro G9 - Glu, Fl - Ser, F2 - His, F3 - h-Phe, F4 - Gly, F - Gly, F6 - Asp, El - Tyr, E2 «. Glu, E3 - Glu, E4 He, E5 - Pro, E6 - Aib, E7 = Aib, E8 - Tyr, E9 - Cha ElO - D-Glu]. Rt- 21.8 min (Sequence Id no. 4). hirunorm-III of structure Ile-Arg-Phe-Thr-Asp-D Ala-Gly-β-Ala-Pro-Glu-Ser-His-h-Phe-Gly-Gly-Asp-Tyr- Glu-Glu-He-Pro-Aib-Aib-P0 4 H 2 -Phe-Cha-D-Glu-OH [compounds of formula (I) wherein Gl = He, G2 = Arg, G3 - Phe, G4 - Thr, G5 = Asp, G6 = D-Ala, G7 - Gly-β Ala, G8 - Pro, G9 - Glu, Fl - Ser, F2 - His, F3 - h Phe, F4 - Gly, F5 - Gly, F6 - Asp, El - Tyr, E2 - Glu E3 - Glu, E4 - He, E5 - Pro, E6 - Aib, E7 - Aib, E8 P0 4 H 2 -Phe, E9 = Cha, ElO - D-Glu], using Boc Phe(P0 4 (CH3)2)-OH.Rt = 20.5 min; chromatographic purit > 99% (Sequence Id no. 5). EXAMPLE 3 The capability of the products described in th present invention of interfering in the coagulatio cascade was evaluated using on human plasma: a) APTT measurements (Babson, A.L. et al., Am. J Clin. Path. 62, 856, (1974), Lenahan, J. G. e al., Clin. Chem., 12, 269, (1966) Miale, J. B Laboratory Medicine: hematology, 6th edition, C.V Mosby Co. (1982) ; b) PT measurements (Quick, A. J. et al., Am. J. Med Sci., 190, 501, (1935), Shapiro, S. et al. Coagulation, Thrombosis Brooklyn Medical Press N.Y. (1949); c) TT measurements (Exner, T. et al., Am. J. Clin Pathol., 71, 521, 1979. The inhibition of the hydrolysis, mediated b thrombin, of Chromozym TH by the peptides of th present invention was evaluated according to th literature (Stone, S.R., Hofsteenge, J. , Biochemistry 25, 4622, (1986). In vitro determination of the peptide resistanc to enzyme hydrolysis by thrombin. 97.2 nmol of hirunorm-I (286 μg) , hirunorm-II (28 μg) , hirunorm-III (293 μg) , were incubated separatel with 0.972 nmol of human a-thrombin (35 μg, 3110 Units NIH/mg) in 800 μL of 0.05 M Tris/HCl buffer, 0.1 M NaCl at pH- 7.8 and a temperature of 37°C. 50 μL Aliquots were diluted with 200 μL of lOOmN H 3 PO and analyzed at pre-set times by capillary electrophoresis on a 0.75 μm x 60 fused silica Waters capillary , using a Waters Quanta 4000 apparatus fitted with an UV detection at 214 nm. The analyses were carried out at a constant voltage of 18 KV, using a lOOmN phosphoric acid buffer for 25 min. The eluted peaks were collected and analyzed by Fab mass spectrometry. In vitro determination of the peptides resistance to hydrolysis by plasma proteases. 1.94 nmol of hirunorm-I (571 μg), hirunorm-II (569 μg) , hirunorm-III (585 μg) , were separately incubated with 1600 μL of human plasma from healthy donors at 37°C for different times. 50 μL aliquots were added with 200 μL of lOOmN H 3 PO 4 . The supernatants were analyzed at pre-set times by HPLC on a Vydac C18 0.46 x 25 cm column with a straight acetonitrile gradient containing 0.1% (v/v) of trifluoroacetic acid (phase B) against 0.1% (v/v) aqueous trifluoroacetic acid (phase A) , from 5 to 70% in B in 35 min at a flow rate of 1 mL/min, with detection at 210 nm. The eluted peaks were collected and analyzed by capillary electrophoresis on a 0.75 μm x 60 cm fused silica Waters capillary, fitted with a UV detector at 214 nm. The analyses were carried out at a constant voltage of 18 KV, using a buffer lOOmN acid phosphoric for 25 min. The eluted peaks were then analyzed by means of Fab mass spectrometry. The inhibition of platelet aggregation thrombin- mediated by the peptides of the present invention was evaluated according to Zucker M.B., Methods in Enzymology, vol. 169, 117-133 and Mustard,J,F, Kinlough-Rathbone, R.L. , Packham, M.A. , Methods in Enzymology, vol.169, 3-11. Comparison of the biological and kinetic parameters of the compounds of the invention with hirudin. Compound PT+ TT+ APTT+ IC 5Q * f t § #§ K^ Hirunorm-I 1.6 0.23 2.7 2.0 >480 300 97. Hirunorm-II 0.32 0.022 0.27 1.0 >480 480 2.6 Hirunorm-III 0.16 0.013 0.18 1.0 >480 480 1.0 Hirunorm-IV >480 2.6 Hirunorm-V -..__— >480 _---»—. 1.0 r-Hirudin$ 1.8 0.086 0.90 + PT, APTT and TT are evaluated as the percent change compared with the control (no inhibitors) and in the table the inhibitor doses (μM) causing a 150%, 250% and 250% increase, respectively, are reported. * ^50 corresponds to the inhibitor doses (nM) causing a 50% decrease of the platelet aggregation. § The factors t fc and t p are the halving times (min) of the inhibitor concentrations in the presence of equimolar amounts of thrombin, respectively both alone and in human plasma. 0 The hydrolysis inhibition constants of a chromogenic substrate are expressed as nM. Hirudin obtained by the recombinant technology. SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: Development Biotechnological Processes snc (B) STREET: Via E.Nicolardi 188 (C) CITY: Napoli (E) COUNTRY: Italy (F) POSTAL CODE (ZIP): 80131 (ii) TITLE OF INVENTION: Thrombine inhibitors their preparation and use for therapeutic prophylactic and diagnostic applications (iii) NUMBER OF SEQUENCES: 5 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Versi #1.25 (EPO) (2) INFORMATION FOR SEQ ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: He Arg Tyr Thr Asp Ala Ala Ala Pro Glu 1 5 10 Asn His Asn Asn Gly Asp 15 Phe Glu Glu He Pro Xaa Xaa Tyr Leu Glu 20 25 Ala in position 6 is D-Ala; Ala in positions 7 and 8 β-Ala; Xaa in positions 22 and 23 is Aib. (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: He Arg Phe Thr Asp Ala Gly Ala Pro Glu 1 5 10 Ser His Phe Gly Gly Asp 15 Tyr Glu Glu He Pro Xaa Xaa Tyr Xaa Glu 20 25 Ala in position 6 is D-Ala; Ala in position 8 is β-Ala Phe in position 13 is h-Phe; Xaa in positions 22 and 2 is Aib; Xaa in position 25 is Cha; Glu in position 2 is D-Glu OH. (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: Xaa Arg Xaa Thr Asp Ala Gly Ala Pro Glu 1 5 10 Ser His Phe Gly Gly Asp 15 Tyr Glu Glu He Pro Xaa Xaa Tyr Xaa Glu 20 25 Xaa in position 1 is Chg; Xaa in position 3 is 2-Na Ala in position 6 is D-Ala; Ala in position 8 is β-Al Phe in position 13 is h-Phe; Xaa in positions 22 and is Aib; Xaa in position 25 is Cha; Glu in position is D-Glu OH. (2) INFORMATION FOR SEQ ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: Xaa Val Xaa Thr Asp Ala Gly Ala Pro Glu 1 5 10 Ser His Phe Gly Gly Asp 15 Tyr Glu Glu He Pro Xaa Xaa Tyr Xaa Glu 20 25 Xaa in position 1 is Chg; Xaa in position 3 is 2-Nal; Ala in position 6 is D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in position 22 and 23 is Aib; Xaa in position 25 is Cha; Glu in position 26 is D-Glu OH. (2) INFORMATION FOR SEQ ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: He Arg Phe Thr Asp Ala Gly Ala Pro Glu 1 5 10 Ser His Phe Gly Gly Asp 15 Tyr Glu Glu He Pro Xaa Xaa Phe Xaa Glu 20 25 Ala in position 6 is D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in positions 22 and 23 is Aib; Phe in position 24 is P0 4 H 2 -Phe; Xaa in position 25 is Cha; Glu in position 26 is D-Glu OH.

Claims: 29 CLAIMS 1. Peptides of general formula (I): G1-G2-G3-G4-G5-G6-G7-G8-G9-F1-F2-F3-F4-F5-F6-E1-E2-E3- E4-E5-E6-E7-E8-E9-E10 wherein: Gl - He, Val, Leu, Cha, Chg, alle, Nle, Pro, Pip, Ph Tyr, Trp, Pgl, 1-Nal, 2-Nal, Met G2 - Arg, Val, L-Pap, L-Map, Ala, He, Leu, Lys, Or Thr, aThr, alle, Ser G3 - 2-Nal, Phe, 1-Nal, Trp, Tyr, Cha, Pgl, Chg G4 - Thr, Ser, Asn, Gin, His, aThr G5 - Asp, Glu G6 - D-Ala, Aib, Gly, Ac g c, Ac r e G7 - -Aca/ ζ -Ava , β -Ala- β -Ala, β -Ala-β-Ac 5 c, β-Ala-β ACgc, β-Ala-Gly, β-Ala-Gaba, Gly-β-Ala, Gly-Gl Gly-Gaba, Gly- -Ava, Gaba-Gly, Gaba-β-Ala, -Ava Gly, Gly-Gly-Gly G8 - Pro, Ala, Gly, ACgC, Ac 5 c G9 - Glu Fl - Ser, Asn, Gin, Thr, aThr, His, h-Ser, h-Phe F2 - His F3 = h-Phe, Leu, Asp, Asn, Gly, Nle, He, alle, Val Cha, Chg, Phe, Tyr, Pgl, Trp, 1-Nal, 2-Nal, Met Deg, Dpg F4 = Gly, Asp, Asn F5 - Gly, Ala, D-Ala, Deg, Dpg F6 - Asp El - Tyr, Phe, Trp, Cha, His, Pgl, 1-Nal, 2-Nal, Chg E2 - Glu E3 - Glu, Pro, Sar, Hyp, ^-Pro, thioproline, Pip, Azt E4 - He, Chg, Cha, E5 - Pro, Sar, Hyp,A-Pro, thioproline, Pip, Azt E6 - Aib, Ala, ACgC, Ac^c, Ac 4 c, AC3C, cis or trans - Glu, (S) or (R)-θi,Me-Glu, Asp, Glu E7 - Aib, Ala, ACgC, Ac^c, AC4C, AC3C, cis or transΔ- Glu, (S) or (R)-fcMe-Glu, Asp, Glu E8 - Tyr, cis or trans Δ-Tyr, (S) or (R)-tyMe-Tyr, Phe, Aib, Ala, para or meta N0 2 -phe, para or met Sθ3H-Phe, para or meta Pθ3H 2 -Phe, para or met P0 4 H 2 -Phe, para or meta S0 4 H-Phe, meta S0 3 H-Tyr, meta P0 3 H 2 -Tyr, para or meta CH 2 Pθ H 2 -Phe, para o meta CH 2 so 3 H-Phe, para or meta CH 2 COOH-Phe, par or meta CO,H-Phe E9 - Cha, Leu, cis or transΔ-Leu, (s) or E10= D-Glu, Glu, β-Ala, Asp, D-Asp, Gly, Ala, Gin, Gla, and salts thereof. 2. Peptides according to claim 1 wherein: Gl - He or Val or Leu or Cha or Chg or alle or Nle or Pro or Phe or Tyr or Trp or Pgl or 1-Nal or 2-Nal or Met; G2 - Arg or L-Pap or L-Map or Val or Thr or He or Ala or Ser; G3 - 2-Nal or Phe or 1-Nal or Trp or Tyr; G4 - Thr; G5 - Asp; G6 - D-Ala; G7 - Gly-β-Ala or β- Ala-β-Ala; G8 - Pro; G9 - Glu, Fl - Ser or Asn or h-Ser or h-Phe; F2 - His; F3 - Nle or Cha or Met or h-Phe; F4 - Gly; F5 = Gly; F6 = Asp; El = Tyr or Phe; E2 = Glu; E3 - Glu or Pro; E4 - He; E5 - Pro; E6 - Aib; E7 - Aib; E8 - Tyr or P0 4 H 2 -Phe; E9 = Cha or Leu; ElO = D- Glu or Glu. 3. Peptides according.to claim 1 wherein: Gl - Chg or He or Val or alle or Nle or Phe or Tyr or Trp or Pgl or 1-Nal or 2-Nal; G2 - Arg or Val ; G3 = 2- Nal or Phe or 1-Nal; G4 - Thr; G5 = Asp; G6 - D-Ala; G7 = Gly-β-Ala; G8 - Pro; G9 - Glu; Fl - Ser; F2 - His; F3 - Nle or h-Phe; F4 = Gly; F5 = Gly; F6 - Asp; El = Tyr or Phe; E2 - Glu; E3 = Glu or Pro; E4 - He; E5 - Pro; E6 = Aib; E7 «- Aib; E8 - Tyr or P0 4 H 2 -Phe; E9 = Cha or Leu; ElO - D-Glu or Glu. 4. Peptide according to claim 1, having Sequence Id no. 1 wherein Ala in position 6 is D-Ala; Ala in positions 7 and 8 is β-Ala; Xaa in positions 22 and 23 is Aib. 5. Peptide according to claim 1, having Sequence Id no. 2 wherein Ala in position 6 is D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in positions 22 and 23 is Aib; Xaa in position 25 is Cha; Glu in position 26 is D-Glu OH. 6. Peptide according to claim 1, having Sequence Id no. 3 wherein Xaa in position 1 is Chg; Xaa in position 3 is 2-Nal; Ala in position 6 id D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in positions 22 and 23 is Aib; Xaa in position 25 is Cha; Glu in position 26 is D-Glu OH. 7. Peptide according to claim 1, having Sequence Id no. 4 wherein Xaa in position 1 is Chg; Xaa in position 3 is 2-Nal; Ala in position 6 is D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in position 22 and 23 is Aib; Xaa in position 25 is Cha; Glu in position 26 is D-Glu OH. 8. Peptide according to claim 1, having Sequence Id no. 5 wherein Ala in. position 6 is D-Ala; Ala in position 8 is β-Ala; Phe in position 13 is h-Phe; Xaa in positions 22 and 23 is Aib; Phe in position 24 is P0 4 H 2 -Phe; Xaa in position 25 is Cha; Glu in positi 26 is D-Glu OH. 9. Pharmaceutical compositions containing o compound of claims 1-8 in combination with a suitab carrier. 10. Compositions according to claim 9, furth comprising a thrombolytic agent. 11. Invasive prostheses coated with one peptide claims 1-8. 12. A diagnostic kit to test the concentration Factor Factor IXa, Factor Xa, thrombin or mixtur thereof in a biological sample, said kit comprising least one peptide of claims 1-8. 13. Peptides of claims 1-8 labelled with radioisotope for ex vivo imaging of fibrin or platel thrombi.

Inventors: DE ROSA ALFREDO, ROSSI ARMANDO, DE ROSA, ALFREDO, ROSSI, ARMANDO
Applicants: DEV BIOTECHNOLOGICAL PROCESSES, ROSA ALFREDO DE, ROSSI ARMANDO, DE ROSA, ALFREDO, DEVELOPMENT BIOTECHNOLOGICAL PROCESSES S.N.C. DI P, ROSSI, ARMANDO
Application year: 1994
Main classification: C07K7
IPC codes: A61K38, A61K51, A61P7, C07K14, G01N33